Spectroscopic identification of a dinuclear metal centre in manganese(II)-activated aminopeptidase P from Escherichia coli: implications for human prolidase

Electron paramagnetic resonance (EPR) spectra and X-ray absorption (EXAFS and XANES) data have been recorded for the manganese enzyme aminopeptidase P (AMPP, PepP protein) from Escherichia coli. The biological function of the protein, a tetramer of 50-kDa subunits, is the hydrolysis of N-terminal Xaa-Pro peptide bonds. Activity assays confirm that the enzyme is activated by treatment with Mn2+. The EPR spectrum of Mn2+-activated AMPP at liquid-He temperature is characteristic of an exchange-coupled dinuclear Mn(II) site, the Mn-Mn separation calculated from the zero-field splitting D of the quintet state being 3.5 (+/- 0.1) Angstrom. In the X-ray absorption spectrum of Mn2+-activated AMPP at the Mn K edge, the near-edge features are consistent with octahedrally coordinated Mn atoms in oxidation state +2. EXAFS data, limited to k less than or equal to 12 Angstrom(-1) by traces of Fe in the protein, are consistent with a single coordination shell occupied predominantly by O donor atoms at an average Mn-ligand distance of 2.15 Angstrom, but the possibility of a mixture of O and N donor atoms is not excluded. The Mn-Mn interaction at 3.5 Angstrom, is not detected in the EXAFS, probably due to destructive interference from light outer-shell atoms. The biological function, amino acid sequence and metal-ion dependence of E. coli AMPP are closely related to those of human prolidase, an enzyme that specifically cleaves Xaa-Pro dipeptides. Mutations that lead to human prolidase deficiency and clinical symptoms have been identified. Several known inhibitors of prolidase also inhibit AMPP. When these inhibitors are added to Mn2+-activated AMPP, the EPR spectrum and EXAFS remain unchanged. It can be inferred that the inhibitors either do not bind directly to the Mn centres, or substitute for existing Mn ligands without a significant change in donor atoms or coordination geometry. The conclusions from the spectroscopic measurements on AMPP have been verified by, and complement, a recent crystal structure analysis.

Solution structure of a defensin-like peptide from platypus venom

Three defensin-like peptides (DLPs) were isolated from platypus venom and sequenced. One of these peptides, DLP-1, was synthesized chemically and its three-dimensional structure was determined using NMR spectroscopy. The main structural elements of this 42-residue peptide were an anti-parallel beta-sheet comprising residues 15-18 and 37-40 and a small 3(10) helix spanning residues 10-12. The overall three-dimensional fold is similar to that of beta-defensin-12, and similar to the sodium-channel neurotoxin ShI (Stichodactyla helianthus neurotoxin I). However, the side chains known to be functionally important in beta-defensin-12 and ShI are not conserved in DLP-1, suggesting that it has a different biological function. Consistent with this contention, we showed that DLP-1 possesses no anti-microbial properties and has no observable activity on rat dorsal-root-ganglion sodium-channel currents.

Structure/function studies of S100A8/A9

The functional importance of members of the S100 Ca2+-binding protein family is recently emerging. A variety of activities, several of which are apparently opposing, are attributed to S100A8, a protein implicated in embryogenesis, growth, differentiation, and immune and inflammatory processes. Murine (m) S100A8 was initially described as a chemoattractant (CP-10) for myeloid cells. It is coordinately expressed with mS100A9 (MRP14) in neutrophils and the non-covalent heterodimer is presumed to be the functional intracellular species. The extracellular chemotactic activity of mS100A8, however, is not dependent on mS100A9 and occurs at concentrations (10(-13)-10(-11) M) at which the non-covalent heterodimer would probably dissociate. This review focuses on the structure and post-translational modifications of mS100A8/A9 and their effects on function, particularly chemotaxis.

Macropore sheath: quantification of plant root and soil macropore association

Plants require roots to supply water, nutrients and oxygen for growth. The spatial distribution of roots in relation to the macropore structure of the soil in which they are growing influences how effective they are at accessing these resources. A method for quantifying root-macropore associations from horizontal soil sections is illustrated using two black vertisols from the Darling Downs, Queensland, Australia. Two-dimensional digital images were obtained of the macropore structure and root distribution for an area 55 x 55 mm at a resolution of 64 mu m. The spatial distribution of roots was quantified over a range of distances using the K-function. In all specimens, roots were shown to be clustered at short distances (1-10 mm) becoming more random at longer distances. Root location in relation to macropores was estimated using the function describing the distance of each root to the nearest macropore. From this function, a summary variable, termed the macropore sheath, was defined. The macropore sheath is the distance from macropores within which 80% of roots are located. Measured root locations were compared to random simulations of root distribution to establish if there was a preferential association between roots and macropores. More roots were found in and around macropores than expected at random.

Bounds on integrals of the Wigner function

The integral of the Wigner function over a subregion of the phase space of a quantum system may be less than zero or greater than one. It is shown that for systems with 1 degree of freedom, the problem of determining the best possible upper and lower bounds on such an integral, over an possible states, reduces to the problem of finding the greatest and least eigenvalues of a Hermitian operator corresponding to the subregion. The problem is solved exactly in the case of an arbitrary elliptical region. These bounds provide checks on experimentally measured quasiprobability distributions.

Visual function: How spiders find the right rock to crawl under

Animals that go on hunting expeditions face the problem of finding the way home at the end of the day. A group of hunting spiders has now been added to the list of animals that use the celestial pattern of polarized light as a compass for navigation. (C) 1999 Elsevier Science Ltd. All rights reserved.

What the evolution of the amyloid protein precursor supergene family tells us about its function

The Alzheimer's disease amyloid protein precursor (APP) gene is part of a multi-gene super-family from which sixteen homologous amyloid precursor-like proteins (APLP) and APP species homologues have been isolated and characterised. Comparison of exon structure (including the uncharacterised APL-1 gene), construction of phylogenetic trees, and analysis of the protein sequence alignment of known homologues of the APP super-family were performed to reconstruct the evolution of the family and to assess the functional significance of conserved protein sequences between homologues. This analysis supports an adhesion function for all members of the APP super family, with specificity determined by those sequences which are not conserved between APLP lineages, and provides evidence for an increasingly complex APP superfamily during evolution. The analysis also suggests that Drosophila APPL and Caenorhabdotids elegans APL-1 may be a fourth APLP lineage indicating that these proteins, while not functional homologues of human APP, are similarly likely to regulate cell adhesion. Furthermore, the beta A4 sequence is highly conserved only in APP orthologues, strongly suggesting this sequence is of significant functional importance in this lineage. (C) 2000 Elsevier Science Ltd. All rights reserved.

Malevolence, stigma, and social distance: Maximizing intergroup differences in HIV/AIDS discourse

This study examined the intergroup language used by young heterosexual Australians in conversations about HIV/AIDS and safe sex. Sixty male and 72 female heterosexuals participated in four-person facilitated conversations (same-sex or mixed-sex) about HIV/AIDS and safe sex, which were recorded and transcribed. We focused on extracts concerning strangers or malevolent individuals who appear to be group members, along with extracts involving foreign national groups. Discourse analysis showed that groups at lower levels of social distance were constructed mainly in terms of individual responsibility. At moderate social distance, stereotypes were more negative, but sub-typing was common, whereas at the highest levels, people were constructed entirely in intergroup terms. The findings of this study suggest that HN prevention programs should make reference to all salient outgroups, so as to neutralize communicative strategies that strengthen intergroup boundaries as a means of reducing perceived personal threat of HIV infection.

An extragalactic HI cloud with no optical counterpart?

We report the discovery, from the H I Parkes All-Sky Survey (HIPASS), of an isolated cloud of neutral hydrogen, which we believe to be extragalactic. The H I mass of the cloud (HIPASS J1712-64) is very low, 1.7 x 10(7) M-circle dot, using an estimated distance of similar to 3.2 Mpc. Most significantly, we have found no optical companion to this object to very faint limits [mu(B) similar to 27 mag arcsec(-2)]. HIPASS J1712-64 appears to be a binary system similar to, but much less massive than, H I 1225 + 01 (the Virgo H. I cloud) and has a size of at least 15 kpc. The mean velocity dispersion measured with the Australia Telescope Compact Array (ATCA) is only 4 km s(-1) for the main component and, because of the weak or nonexistent star formation, possibly reflects the thermal line width (T < 2000 K) rather than bulk motion or turbulence. The peak column density for HIPASS J1712-64, from the combined Parkes and ATCA data, is only 3.5 x 1019 cm(-2), which is estimated to be a factor of 2 below the critical threshold for star formation. Apart from its significantly higher velocity, the properties of HIPASS J1712-64 are similar to the recently recognized class of compact high-velocity clouds. We therefore consider the evidence for a Local Group or Galactic origin, although a more plausible alternative is that HIPASS J1712-64 was ejected from the interacting Magellanic Cloud-Galaxy system at perigalacticon similar to 2 x 10(8) yr ago.

Long-distance signalling and a mutational analysis of branching in pea

Four ramosus mutants with increased branching at basal and aerial nodes have been used to investigate the genetic regulation of bud outgrowth in Pisum sativum L. (garden pea). Studies of long-distance signalling, xylem sap cytokinin concentrations, shoot auxin level, auxin transport and auxin response are discussed. A model of branching control is presented that encompasses two graft-transmissible signals in addition to auxin and cytokinin. Mutants rms1 through rms4 are not deficient in indole-3-acetic acid (IAA) or in the basipetal transport of this hormone. Three of the four mutants, rms1, rms3 and rms4, have very reduced cytokinin concentrations in xylem sap from roots. This reduction in xylem sap cytokinin concentration appears to be caused by a property of the shoot and may be part of a feedback mechanism induced by an aspect of bud outgrowth. The shoot-to-root feedback signal is unlikely to be auxin itself, as auxin levels and transport are not correlated with xylem sap cytokinin concentrations in various intact and grafted mutant and wild-type plants. Rms1 and Rms2 act in shoot and rootstock to regulate the level or transport of graft-transmissible signals. Various grafting studies and double mutant analyses have associated Rms2 with the regulation of the shoot-to-root feedback signal. Rms1 is associated with a second unknown graft-transmissible signal that is postulated to move in the direction of root-to-shoot. Exogenous auxin appears to interact with both of the signals regulated by Rms1 and Rms2 in the inhibition of branching after decapitation. The action of Rms3 and Rms4 is less apparent at this stage, although both appear to act largely in the shoot.

Vehicle and driver attributes affecting distance from the steering wheel in motor vehicles

The current study was designed to confirm that female drivers sit closer to the steering wheel than do male drivers and to investigate whether this expected difference in sitting position is attributable to differences in the physical dimensions of men and women. Driver body dimensions and multiple measures of sitting distance from the steering wheel were collected from a sample of 150 men and 150 women. The results confirmed that on average, women sit closer to the steering wheel than men do and that this difference is accounted for by variations in body dimensions, especially height. This result suggests that driver height may provide a good surrogate for sitting distance from the steering wheel when investigating the role of driver position in real-world crash outcomes. The potential applications of this research include change to vehicle design that allows independent adjustment of the relative distance among the driver's seat, the steering wheel, and the floor pedals.